Electrogeneration of a Free-Standing Cytochrome c-Silica Matrix at a Soft Electrified Interface.
Electrogeneration of a Free-Standing Cytochrome c-Silica Matrix at a Soft Electrified Interface.
复制标题
DOI：
10.1021/acs.langmuir.1c00409
复制
发表时间：
2021-04-06
期刊：
影响因子：
--
通讯作者：
Herzog G
中科院分区：
文献类型：
--
作者：
Gamero-Quijano A;Dossot M;Walcarius A;Scanlon MD;Herzog G
蛋白质与固-液或液-液界面的相互作用可能会破坏其构象的稳定性，从而导致生物活性的丧失。我们在这里提出了一种在带电的液-液界面上固定化蛋白质的方法。细胞色素c(Cytc)在带电的液-液界面处通过电化学过程被包裹在二氧化硅基质中。二氧化硅的缩合是由界面电位窗口下端的阳离子表面活性剂十六烷基三甲基铵的界面转移引发的。然后，当界面电位Δowϕ处于电位窗口的正端时，Cytc被吸附在先前电沉积的二氧化硅层上。原位UV-Vis吸收光谱表明，通过电位窗口的来回循环，二氧化硅的电沉积和Cytc的吸附依次发生。从液-液界面收集后，用紫外可见漫反射光谱、共聚焦拉曼显微镜和荧光显微镜对Cytc-SiO_2基质进行了异位表征，表明该蛋白质在包裹过程中保持了其三级结构。通过对O2没有电催化活性(在Cytc变性的情况下观察到)，进一步证实了没有变性。这种蛋白质包埋方法也可用于开发两相生物电合成或生物电催化应用中的其他蛋白质(如铁-S簇状氧化还原酶、含铜还原酶、含吡咯喹啉醌的酶或黄素蛋白)。
Interactions of a protein with a solid–liquid or a liquid–liquid interface may destabilize its conformation and hence result in a loss of biological activity. We propose here a method for the immobilization of proteins at an electrified liquid–liquid interface. Cytochrome c (Cyt c) is encapsulated in a silica matrix through an electrochemical process at an electrified liquid–liquid interface. Silica condensation is triggered by the interfacial transfer of cationic surfactant, cetyltrimethylammonium, at the lower end of the interfacial potential window. Cyt c is then adsorbed on the previously electrodeposited silica layer, when the interfacial potential, Δowϕ, is at the positive end of the potential window. By cycling of the potential window back and forth, silica electrodeposition and Cyt c adsorption occur sequentially as demonstrated by in situ UV–vis absorbance spectroscopy. After collection from the liquid–liquid interface, the Cyt c–silica matrix is characterized ex situ by UV–vis diffuse reflectance spectroscopy, confocal Raman microscopy, and fluorescence microscopy, showing that the protein maintained its tertiary structure during the encapsulation process. The absence of denaturation is further confirmed in situ by the absence of electrocatalytic activity toward O2 (observed in the case of Cyt c denaturation). This method of protein encapsulation may be used for other proteins (e.g., Fe–S cluster oxidoreductases, copper-containing reductases, pyrroloquinoline quinone-containing enzymes, or flavoproteins) in the development of biphasic bioelectrosynthesis or bioelectrocatalysis applications.
登录
查看更多内容
影响因子：
3.3
作者：
Cheng, Shih-Hsun;Kao, Kun-Che;Lee, Chia-Hung
通讯作者：
Lee, Chia-Hung
影响因子：
7.4
作者：
Felisilda, Bren Mark B.;Arrigan, Damien W. M.
通讯作者：
Arrigan, Damien W. M.
影响因子：
3.9
作者：
Haldar, Shubhasis;Sil, Pallabi;Chattopadhyay, Krishnananda
通讯作者：
Chattopadhyay, Krishnananda
影响因子：
7.4
作者：
Hartvig, Rune A.;Mendez, Manuel A.;Jensen, Henrik
通讯作者：
Jensen, Henrik
影响因子：
3.9
作者：
Gamero-Quijano, Alonso;Huerta, Francisco;Montilla, Francisco
通讯作者：
Montilla, Francisco